The Molecular Biology Core will serve two main functions for the Gene Therapy Center. First, it will provide all the necessary molecular biology reagents for the isolation of genes, recombinant DNA construction, vector construction and preparation. Secondly, it will provide in situ hybridization techniques to detect the successful transfer of genes and their expression. The service of ligonucleotide synthesis and preparation of viral vectors will be provided. In addition, the Core will coordinate automatic DNA synthesis, peptide synthesis, protein microsequencing, FACS analysis, and cell sorting. It will also provide training and advice for the strategies of gene cloning and mutagenesis. This core will also provide in situ hybridization on cells transfected by various.gene transfer vehicles, to detect DNA expression and immunocytochemistry to detect the protein products including CFTR. These studies will be useful for the determination of the appropriate promoter for tissue specific expression as well as for the detection of the level of expression of the gene. The staff of this Core will be strongly committed to the continuous introduction of new technology and will train investigators, postdoctoral fellows, and laboratory technicians in the techniques.